Compositions for the delivery of therapeutic agents and uses thereof

Abstract

The present invention provides drug delivery vehicles comprising polytheylyene-lipid conjugates (PEG-lipid), wherein the circulation lifetime and biodistribution of the drug delivery vehicles are regulated by the PEG-lipid. More particularly, the present invention provides liposomes, SNALP and SPLP comprising such PEG-lipid conjugates, and methods of using such compositions to selectively target a tumor site or other tissue of interest (e.g., liver, lung, spleen, etc.).

1 . A method of introducing a nucleic acid into a tumor cell, said method comprising contacting said tumor cell with a nucleic acid-lipid particle comprising a cationic lipid, a noncationic lipid, a PEG-lipid conjugate, and a nucleic acid, wherein the alkyl or acyl chains of the lipid portion of said PEG-lipid conjugate comprise from 12 to 20 carbon atoms.  
   
   
       2 . The method in accordance with  claim 1 , wherein said cationic lipid is a member selected from the group consisting of: 1,2-DiLinoleyloxy-N,N-dimethylaminopropane (DLinDMA), 1,2-Dilinolenyloxy-N,N-dimethylaminopropane (DLenDMA), N,N-dioleyl-N,N-dimethylammonium chloride (DODAC), N,N-distearyl-N,N-dimethylammonium bromide (DDAB), N-(1-(2,3-dioleoyloxy)propyl)-N,N,N-trimethylammonium chloride (DOTAP), N-(1 -(2,3-dioleyloxy)propyl)-N,N,N-trimethylammonium chloride (DOTMA), and N,N-dimethyl-2,3-dioleyloxy)propylamine (DODMA), and a mixture thereof.  
   
   
       3 . The method in accordance with  claim 1 , wherein said noncationic lipid is a member selected from the group consisting of: (DSPC), dioleoylphosphatidylcholine (DOPC), dipalmitoylphosphatidylcholine (DPPC), dioleoylphosphatidylglycerol (DOPG), dipalmitoylphosphatidylglycerol (DPPG), dioleoyl-phosphatidylethanolamine (DOPE), palmitoyloleoylphosphatidylcholine (POPC), palmitoyloleoyl- phosphatidylethanolamine (POPE) and dioleoyl- phosphatidylethanolamine 4-(N-maleimidomethyl)-cyclohexane-1-carboxylate (DOPE-mal), dipalmitoyl phosphatidyl ethanolamine (DPPE), dimyristoylphosphoethanolamine (DMPE), distearoyl-phosphatidyl-ethanolamine (DSPE), 16-O-monomethyl PE, 16-O-dimethyl PE, 18-1-trans PE, 1-stearoyl-2-oleoyl-phosphatidyethanolamine (SOPE), and 1,2-dielaidoyl-sn-glycero-3-phophoethanolamine (transDOPE).  
   
   
       4 . The method in accordance with  claim 1 , wherein the alkyl or acyl chains of the lipid portion of said PEG-lipid conjugate comprise from 16 to 20 carbon atoms.  
   
   
       5 . The method in accordance with  claim 1 , wherein said noncationic lipid is an anionic lipid.  
   
   
       6 . The method in accordance with  claim 1 , wherein said noncationic lipid is a neutral lipid.  
   
   
       7 . The method in accordance with  claim 1 , wherein said PEG-lipid is a member selected from the group consisting of a PEG-diacylglycerol (DAG), a PEG dialkyloxypropyl (DAA), a PEG-phospholipid, a PEG-ceramide, and combinations thereof.  
   
   
       8 . The method in accordance with  claim 1 , wherein said PEG-lipid is a PEG dialkyloxypropyl (DAA) selected from the group consisting of: a PEG-dipalmityloxypropyl (C 16 ); a PEG-distearyloxypropyl (C 18 ); and a PEG-diicosyloxypropyl (C 20 ).  
   
   
       9 . The method in accordance with  claim 1 , wherein said PEG-lipid is PEG-dialkyloxypropyl (DAA) having the following structure:  
     
       
         
         
             
             
         
       
     
     wherein: 
 R 1  and R 2  are independently selected and are alkyl groups having from about 16 to about 20 carbon atoms;  
 PEG is a polyethyleneglycol; and  
 L is a non-ester containing linker moiety.  
 
   
   
       10 . The method in accordance with  claim 1 , wherein said PEG-lipid is PEG-diacylglycerol (DAG) selected from the group consisting of a PEG-dipalmitoylglycerol (C 16 ), a PEG-disterylglycerol (C 18 ) and a PEG-diicosylglycerol (C20).  
   
   
       11 . The method in accordance with  claim 1 , wherein said PEG-lipid is PEG-ceramide (Cer) selected from the group consisting of PEG-ceramide (C 16 ), a PEG-ceramide (C 18 ) and PEG-ceramide (C 20 ).  
   
   
       12 . The method in accordance with  claim 1 , wherein said cationic lipid comprises from about 5% to about 45% of the total lipid present in said particle.  
   
   
       13 . The method in accordance with  claim 1 , wherein said cationic lipid comprises from about 5% to about 15% of the total lipid present in said particle.  
   
   
       14 . The method in accordance with  claim 1 , wherein said cationic lipid comprises from about 30% to about 50% of the total lipid present in said particle.  
   
   
       15 . The method in accordance with  claim 1 , wherein said cationic lipid comprises about 40% of the total lipid present in said particle.  
   
   
       16 . The method in accordance with  claim 1 , wherein said noncationic lipid comprises from about 5% to about 90% of the total lipid present in said particle.  
   
   
       17 . The method in accordance with  claim 1 , wherein said noncationic lipid comprises from about 20% to about 85% of the total lipid present in said particle.  
   
   
       18 . The method in accordance with  claim 1 , wherein said PEG-lipid conjugate comprises from 1% to about 20% of the total lipid present in said particle.  
   
   
       19 . The method in accordance with  claim 1 , wherein said PEG-lipid conjugate comprises from 2% to about 15% of the total lipid present in said particle.  
   
   
       20 . The nucleic acid-lipid particle in accordance with  claim 1 , wherein said PEG-lipid conjugate comprises about 2% of the total lipid present in said particle.  
   
   
       21 . The method in accordance with  claim 1 , wherein said noncationic lipid is DSPC.  
   
   
       22 . The method in accordance with  claim 1 , wherein said nucleic acid-lipid particle further comprises cholesterol.  
   
   
       23 . The method in accordance with  claim 22 , wherein the cholesterol comprises from about 0% to about 10% of the total lipid present in said particle.  
   
   
       24 . The method in accordance with  claim 22 , wherein the cholesterol comprises from about 10% to about 60% of the total lipid present in said particle.  
   
   
       25 . The method in accordance with  claim 22 , wherein the cholesterol comprises from about 20% to about 45% of the total lipid present in said particle.  
   
   
       26 . The method in accordance with  claim 1 , wherein said nucleic acid is DNA.  
   
   
       27 . The method in accordance with  claim 1 , wherein said nucleic acid is a plasmid.  
   
   
       28 . The method in accordance with  claim 1 , wherein said nucleic acid is an antisense oligonucleotide.  
   
   
       29 . The method in accordance with  claim 1 , wherein said nucleic acid is a ribozyme.  
   
   
       30 . The method in accordance with  claim 1 , wherein said nucleic acid is a small interfering RNA (siRNA).  
   
   
       31 . The method in accordance with  claim 1 , wherein said nucleic acid encodes a therapeutic product of interest.  
   
   
       32 . The method in accordance with  claim 31 , wherein said therapeutic product of interest is a peptide or protein.  
   
   
       33 . The method in accordance with  claim 31 , wherein said therapeutic product of interest is a small interfering RNA (siRNA).  
   
   
       34 . The method in accordance with  claim 1 , wherein the nucleic acid in said nucleic acid-lipid particle is not substantially degraded after exposure of said particle to a nuclease at 37° C. for 20 minutes.  
   
   
       35 . The method in accordance with  claim 1 , wherein the nucleic acid in said nucleic acid-lipid particle is not substantially degraded after incubation of said particle in serum at 37° C. for 30 minutes.  
   
   
       36 . The method in accordance with  claim 1 , wherein the nucleic acid is fully encapsulated in said nucleic acid-lipid particle.  
   
   
       37 . A method of introducing a nucleic acid to the lung of a mammal, said method comprising administering to said mammal a nucleic acid-lipid particle comprising a cationic lipid, a noncationic lipid, a PEG-lipid conjugate, and a nucleic acid, wherein the alkyl or acyl chains of the lipid portion of said PEG-lipid conjugate comprise from 16 to 20 carbon atoms.  
   
   
       38 . The method in accordance with  claim 37 , wherein said PEG-lipid is a PEG dialkyloxypropyl (DAA) slected from the group consisting of a PEG-dipalmityloxypropyl (C 16 ), PEG-distearyloxypropyl (C 18 ), and a PEG-diicosyloxypropyl (C 20 ).  
   
   
       39 . The method in accordance with  claim 37 , wherein said PEG-lipid is PEG-diacylglycerol (DAG) selected from the group consisting of: a PEG-dipalmitoylglycerol (C 16 ), and a PEG-disterylglycerol (C 18 ).  
   
   
       40 . The method in accordance with  claim 37 , wherein said PEG-lipid is PEG-ceramide (Cer) selected from the group consisting of: PEG-ceramide (C 16 ), and a PEG-ceramide (C 18 ) and PEG-ceramide (C 20 ).  
   
   
       41 . A method of introducing a nucleic acid to the liver of a mammal, said method comprising administering to said mammal a nucleic acid-lipid particle comprising a cationic lipid, a noncationic lipid, a PEG-lipid conjugate, and a nucleic acid, wherein the alkyl or acyl chains of the lipid portion of said PEG-lipid conjugate comprise from 8 to 14 carbon atoms.  
   
   
       42 . The method in accordance with  claim 41 , wherein said PEG-lipid is as PEG dialkyloxypropyl (DAA) slected from the group consisting of a PEG-dilauryloxypropyl (C 12 ) and a PEG-dimyristyloxypropyl (C 14 ).  
   
   
       43 . The method in accordance with  claim 41 , wherein said PEG-lipid is PEG-diacylglycerol (DAG) selected from the group consisting of: PEG-dilaurylglycerol (C 12 ) and a PEG-dimyristylglycerol (C 14 ).  
   
   
       44 . The method in accordance with  claim 41 , wherein said PEG-lipid is PEG-ceramide (Cer) selected from the group consisting of: PEG-ceramide (C 12 ) and a PEG-ceramide (C 14 ).  
   
   
       45 . A method of introducing a nucleic acid to the spleen of a mammal, said method comprising administering to said mammal a nucleic acid-lipid particle comprising a cationic lipid, a noncationic lipid, a PEG-lipid conjugate, and a nucleic acid, wherein the alkyl or acyl chains of the lipid portion of said PEG-lipid conjugate comprise from 8 to 14 carbon atoms.  
   
   
       46 . The method in accordance with  claim 45 , wherein said PEG-lipid is a PEG dialkyloxypropyl (DAA) slected from the group consisting of a PEG-dilauryloxypropyl (C 12 ) and a PEG-dimyristyloxypropyl (C 14 ).  
   
   
       47 . The method in accordance with  claim 45 , wherein said PEG-lipid is PEG-diacylglycerol (DAG) selected from the group consisting of: PEG-dilaurylglycerol (C 12 ) and a PEG-dimyristylglycerol (C 14 ).  
   
   
       48 . The method in accordance with  claim 45 , wherein said PEG-lipid is PEG-ceramide (Cer) selected from the group consisting of selected from the group consisting of PEG-ceramide (C 12 ) and a PEG-ceramide (C 14 ).